Hydrophobic Interaction
Chromatography Media

CellufinecTM MAX Butyl
CellufineTM MAX Phenyl

CellufineTM MAX is a 2nd generation Cellufine media with high flow characteristics. JNC developed a new, highly cross-linked base resin for Cellufine MAX series. Cellufine MAX hydrophobic interaction chromatography is now available with MAX Phenyl and Butyl chemistries.

SEM analysis of Cellufine MAX base resin
SEM analysis of Cellufine MAX base resin

Cellulose, a natural polysaccharide, possesses unique crystalline molecular structure differing from non-crystalline polysaccharides such as agarose. Thus Cellufine has distinctive pore structure as shown in the pictograph (Fig. 1). The new Cellufine MAX series offers the largest pore size of all Cellufine chromatography media. The benefit of such pore size in Cellufine MAX HIC media provides superior strength and excellent mass transfer.

Partial structure

  • Hydrophobic Interaction

    Cellufine MAX Butyl

    Cellufine MAX Butyl
  • Cellufine MAX Phenyl
    Cellufine MAX Phenyl LS

    Cellufine MAX Phenyl,Cellufine MAX Phenyl LS

Characteristics

Characteristics
Type Cellufine MAX Butyl Cellufine MAX Phenyl Cellufine MAX Phenyl LS
Matrix Highly Cross-linked Cellulose
Particle size 40~130 μm
Ligand type Butyl Phenyl
BSA adsorption capacity (mg/ml) ≧9 ≧11 ≧4
BSA elution efficiency (%) > 70 > 35 > 65
Polyclonal IgG 10% DBC (mg/ml) 17 30 19
Operating pressure < 0.3 MPa
pH stability pH 2 ~ 13
Storage 20 % Ethanol

Model Protein Separation

Model Protein Separation
Column
6.6 mm ID x 50 mm L
Buffer A
10 mM phosphate buffer (pH 7) + 1.5 M (NH4)2SO4
Buffer B
10 mM phosphate (pH 7)
Proteins
Ribonuclease A, Lysozyme, α-Chymotripsinogen A

Adsorption Capacity of Model Proteins

Adsorption Capacity of Model Proteins
Column
5 mm ID x 10 cm L
Protein concentration
1 mg/ml
Buffer
50 mM Tris-HCl (pH 8.5)

Dynamic Binding Capacity for Cellufine MAX Phenyl vs. Salt Concentration

Dynamic Binding Capacity for Cellufine MAX Phenyl vs. Salt Concentration
Column
5 mm I.D. x 5 cm L
Flow velocity
150 cm/hr
Protein concentration
1 mg/ml
Buffer
20 mM Phosphate (pH7.0) + (NH4)2SO4

Dynamic Adsorption Capacity for Cellufine MAX Butyl

Dynamic Adsorption Capacity for Cellufine MAX Butyl
Column
5 mm I.D. x 5 cm L
Flow rate
0.5 ml/ min
Protein concentration
1 mg/ml
Buffer
10 mM Phosphate (pH 7.0) +
2 M (NH4)2SO4 / BSA
1 M (NH4)2SO4 / polyclonal IgG

Pressure vs. Flow Properties

Pressure vs. Flow Properties
Column
2.2cm I.D. x 20 cm L
Temperature
24 ± 1 ℃
Mobile phase
Water

Ligand density is controllable in Cellufine MAX HIC Ex.) Purification of HBsAg by Cellufine MAX Butyl

As ligand density is controllable, Cellufine MAX HIC is optimized for customer’s purpose.
The figure shows a chromatogram of adsorption-desorption of r Hepatitis B virus particles to Cellufine MAX Butyl LS (optimization for HBV) and Agarose typed butyl media

Ligand density is controllable in Cellufine MAX HIC Ex.) Purification of HBsAg by Cellufine MAX Butyl
Cellufine MAX Butyl LS Agarose typed Butyl media
Load 163 (100) 163 (100)
Flowthrough 48 (29) 50 (31)
Eluted target 78 (48) 63 (39)

μg(%)

Run condition

HIC resin
Cellufine MAX Butyl LS and Agarose typed butyl media
System
AKTA explorer 10S
Column
Φ3.0 x 50 mm (vol. 0.35 ml)
Sample
150 mg of r-HBsAg (Beacle Inc.) in 0.02 M PB, 0.6M (NH4)2SO4, pH 7.0
Equilibration and wash
0.02 M PB, 0.6M (NH4)2SO4, pH 7.0
Elution 1
0.02 M PB, pH 7.0
Flow rate
0.25 ml/min(220 cm/hr)

Chemical and Caustic Stability

  • Ethanol (70%)
  • NaOH (0.5M)
  • Isopropyl alcohol (30%)
  • Detergents
  • Guanidine hydrochloride (6M)
  • Autoclave (121 ºC, 20 min)
  • Urea (6M)

Development of the grade

Development of the grade according to demand of a customer (inquiry)

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